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Limbal microenvironment can induce transdifferentiation of hair follicle stem cells into corneal epithelial‐like cells
Author(s) -
BLAZEJEWSKA EA,
SCHLOETZERSCHREHARDT U,
KRUSE FE
Publication year - 2008
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2008.4233.x
Subject(s) - microbiology and biotechnology , stem cell , laminin , fibronectin , biology , transdifferentiation , hair follicle , stromal cell , extracellular matrix , basement membrane , cellular differentiation , myofibroblast , pathology , fibrosis , cancer research , medicine , biochemistry , gene
Abstract Purpose To investigate the plasticity of murine vibrissa hair follicle (HF) stem cells regarding differentiation into corneal epithelial‐like cells through modulation by limbus‐specific microenvironmental factors. Methods HFs were isolated and the dissociated bulge stem cells enriched by clonal expansion, and subcultivated on various extracellular matrices (collagen type IV, laminin‐1, laminin‐5, fibronectin) in DMEM/F12 medium supplemented with different conditioned media (CMs). CMs were derived from central and peripheral corneal fibroblasts and limbal stromal fibroblasts. Growth potential and cellular phenotype were evaluated by light and electron microscopy, real‐time‐PCR and immunohistochemistry using antibodies against stem cell (CK15, integrin α6) and differentiation markers (CK12) or (CK10). Results Laminin‐5 and collagen IV promoted rapid cell adhesion, proliferation, and generation of confluent, regularly arranged epitheloid cell sheets, whereas laminin‐1 and fibronectin adversely affected these cell properties. Addition of CMs differentially influenced cellular phenotype and differentiation. Limbal fibroblast CM induced differentiation of cuboid cells showing ultrastructural characteristics of a corneal epithelial phenotype. Moreover, it markedly increased the number of CK12 immunopositive cells while decreasing expression of CK10 opposed to the other CMs. Conclusion Hair follicle stem cells are capable of differentiating into corneal epithelial‐like cells in vitro when exposed to a limbus‐specific microenvironment. Therefore, the HF may be an alternative therapeutic source of multipotent stem cells for generation of autologous epithelial cell sheets for ocular surface reconstruction.